Project Summary: Alcohol use disorder (AUD) can be characterized by a pattern of compulsive alcohol drinking or loss of control over alcohol drinking. The positive and negative effects of ethanol appear to be regulated by genetic and epigenetic changes in several key brain regions. Long-term alcohol use causes structural and functional changes in the prefrontal cortex (PFC), hippocampus, amygdala, and ventral tegmental area of the brain which may drive behavioral phenotypes, such as anxiety, depression, motivation, and increased alcohol drinking. Several protein families such as histone deacetylases (HDACs), histone acetyltransferases (HATs), histone methyltransferases (HMTs), DNA methyltransferases (DNMTs), lysine demethylases, and DNA demethylases are important players in chromatin remodeling within the genome and are altered by chronic ethanol exposure and withdrawal. However, the interplay between these different epigenetic modifiers lead to an altered state of the epigenome and their role in transcriptomic changes associated with adaptations in the reward and stress systems in the brain (VTA, amygdala, PFC, and hippocampus) during AUD is still unclear. The overall aim of this Alcohol Research Center is to evaluate the epigenetic and genetic basis of molecular changes in the brain that underlie behavioral changes in AUD. This P50 application entitled ?Center for Alcohol Research in Epigenetics (CARE)? consists of four highly inter-related preclinical and translational research projects [research project #1, VTA, (Brodie /Glover), research project #2, Amygdala, (Pandey), research project #3, Hippocampus, (Lasek) and research project #4, PFC, (Guidotti/Grayson/Gavin)] and two current pilot projects [pilot project #1 (Epigenetic biomarkers in AUD), and pilot project #2 (Epitranscriptomic modifications in AUD)], and three Cores [Administrative (Pandey/Lasek), Epigenetics (Grayson/Maienschein-Cline), and Behavioral Core (Glover/Zhang)]. In addition, the CARE will serve as an important resource and provide a scientifically enriched environment for training opportunities for the next generation of neuroscientists studying AUD and will disseminate scientific knowledge of AUD to the general public through a community outreach program. The primary thematic focus of CARE, as a whole, will be to mechanistically link emerging novel molecular targets identified by whole-genome approaches (ATAC-seq, ChIP-seq, RNA-seq, and DNA methylation/demethylation EPIC array) in the proposed brain circuitry to behavioral phenotypes. Mechanistic approaches (CRISPR-dCas9, shRNA, siRNA, chemogenetic) will be used to manipulate the epigenome through alterations of either histone acetylation/methylation mechanisms or DNA methylation mechanisms, in key brain circuitry that regulates behavioral phenotypes associated with AUD (anxiety, depression, motivation, and alcohol drinking) in order to better understand the pathophysiology of AUD and develop better pharmacotherapy.